The short-term goals of this application are to identify adducted and altered protein biomarkers that correlate[unreadable] with organophosphate (OP) insecticide exposure and to use these biomarkers to generate antibodies to[unreadable] validate the detection of these biomarkers in biological fluids. Over 100 million pounds of OP insecticides[unreadable] are used each year but lead to over 25,000 reports to poison control centers each year including 10,000[unreadable] involving children under age six. These reports only account for acute poisoning events and do not include[unreadable] low dose or chronic exposures. In addition to general exposure and food contamination, the safety of OPs is[unreadable] a large public health concern because OPs share chemical traits, structure and biological mechanism with[unreadable] nerve gas agents. These concerns are elevated by an array of neurologic and non-neurologic sequelae that[unreadable] have been reported in connection with exposure to OPs. To combat, monitor and provide therapeutic[unreadable] intervention, a blood cholinesterase test (BCT) has been conducted for decades. However, the test is limited[unreadable] and inadequate to assess OP exposure and its shortcomings identified by an EPA report questioned the[unreadable] merit of the BCT and suggested the need to examine true biomarkers of exposure. At this time, there is a[unreadable] serious void in effective tests of OP exposure. New tests are needed that are sensitive, selective, operate in[unreadable] real time and are based on reliable, well-characterized OP-biomarkers derived from exacting molecular[unreadable] events that correlate with cellular, tissue, organ or systemic toxicity.[unreadable] The long range goal of our research is to identify OP-adducted and/or OP-altered protein biomarkers that are[unreadable] associated with OP-induced sequelae and to develop diagnostic methods and tests that assess the human[unreadable] health risk and aid therapeutic action. For the proposed grant period, we will address the following specific[unreadable] aims: SA 1. Show that OP's result in highly specific OP-AChE adducts that can be identified and[unreadable] differentiated by antibodies. SA2. Identify and characterize OP-adducted protein biomarkers in SH-SY5Y[unreadable] neuroblastoma cells using OP-reporter molecules. SA3. Identify and characterize OP-protein biomarkers[unreadable] from human saliva, human serum and human erythrocytes and SA4. Prepare customized antibodies that[unreadable] recognize OP-adducted proteins identified in SA-II and SA-III and develop efficient diagnostic tests to identify[unreadable] and quantify OP-protein adducts.